Auxin efflux by PIN-FORMED proteins is activated by two different protein kinases, D6 PROTEIN KINASE and PINOID.
The development and morphology of vascular plants is critically determined by synthesis and proper distribution of the phytohormone auxin. The directed cell-to-cell distribution of auxin is achieved through a system of auxin influx and efflux transporters. PIN-FORMED (PIN) proteins are proposed auxin efflux transporters, and auxin fluxes can seemingly be predicted based on the--in many cells--asymmetric plasma membrane distribution of PINs. Here, we show in a heterologous Xenopus oocyte system as well as in Arabidopsis thaliana inflorescence stems that PIN-mediated auxin transport is directly activated by D6 PROTEIN KINASE (D6PK) and PINOID (PID)/WAG kinases of the Arabidopsis AGCVIII kinase family. At the same time, we reveal that D6PKs and PID have differential phosphosite preferences. Our study suggests that PIN activation by protein kinases is a crucial component of auxin transport control that must be taken into account to understand auxin distribution within the plant.


Localization-dependent translation requires a functional interaction between the 5' and 3' ends of oskar mRNA.
The precise restriction of proteins to specific domains within a cell plays an important role in early development and differentiation. An efficient way to localize and concentrate proteins is by localization of mRNA in a translationally repressed state, followed by activation of translation when the mRNA reaches its destination. A central issue is how localized mRNAs are derepressed. In this study we demonstrate that, when oskar mRNA reaches the posterior pole of the Drosophila oocyte, its translation is derepressed by an active process that requires a specific element in the 5' region of the mRNA. We demonstrate that this novel type of element is a translational derepressor element, whose functional interaction with the previously identified repressor region in the oskar 3' UTR is required for activation of oskar mRNA translation at the posterior pole. The derepressor element only functions at the posterior pole, suggesting that a locally restricted interaction between trans-acting factors and the derepressor element may be the link between mRNA localization and translational activation. We also show specific interaction of two proteins with the oskar mRNA 5' region; one of these also recognizes the 3' repressor element. We discuss the possible involvement of these factors as well as known genes in the process of localization-dependent translation.


The effect of shade, shearing and wool type in the protection of Merino sheep from Hypericum perforatum (St John's wort) poisoning.
To investigate the roles of shade, fleece length and wool type in the protection of sheep from Hypericum perforatum poisoning. Adult Merino ewes of superfine, fine and medium wool type. Seventy sheep were divided into seven equal groups. During late spring and summer a series of successive, replicate experiments was conducted, each using one group and lasting 5 days. The sheep carried 14 to 24 weeks wool growth. In each experiment the treatments tested were Hypericum +, sunlight + (n = 7); Hypericum +, sun - (n = 1); Hypericum -, sun + (n = 1); Hypericum -, sun - (n = 1). Next, 24 sheep in two equal groups were used in experiments of similar design to the above. Each group consisted of nine recently (1 to 3 weeks previously) shorn and three wool covered (25 to 26 weeks growth) sheep. The treatments tested were Hypericum +, sunlight +, fleece - (n = 9); Hypericum +, sun -, fleece + (n = 1); Hypericum -, sun +, fleece + (n = 1); Hypericum -, sun -, fleece + (n = 1). Finely milled Hypericum was administered by gavage to provide 3 mg hypericin / kg body weight. Sheep were sheltered from direct sunlight or were exposed for 5 h per day for 4 successive post-treatment days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C was considered indicative of hypericin poisoning. After Hypericum treatment hypericin poisoning was displayed by 26.5% of woolled sheep that were exposed to sunlight, but by none of those that were fully shaded. In similarly treated but recently shorn sheep 94% displayed hypericin poisoning when exposed to sunlight. In the wool covered group the percentages of poisoned animals based on wool type were: superfine 14%, fine 28.5%, medium 33.3%. In the recently shorn group the percentage for all three approached 100%. A majority of Merinos with at least 14 weeks wool growth will not be poisoned by a single oral dose of 3 mg hypericin /kg, but because hypericin persists in the blood circulation for several days this safe dose will be lowered by continuous daily ingestion. Sheep with access to substantial areas of shade could safely ingest much greater amounts of hypericin. Wool removal greatly increases the risk of poisoning. Superfine Merinos with a wool cover should be able to ingest more hypericin than comparable, medium wool types, without any increased risk of poisoning. The ability of ruminant livestock to safely ingest Hypericum is probably determined more by the amount of skin protection they have against incident sunlight than by differences in hypericin metabolism and excretion capacity.


Toward a new communications framework for HIV/AIDS.
In response to the overwhelming burden of new cases of human immunodeficiency virus (HIV) in Africa, Asia, Latin America, and the Caribbean, the Joint United Nations Programme on HIV/AIDS (UNAIDS), in 1997, initiated a project to examine the application of existing communication theories/models to HIV/acquired immune deficiency virus (AIDS) prevention and care in these regions. In the past 2 years, 103 leading researchers and practitioners from different parts of the world were invited by the UNAIDS to participate in one of five consultative workshops designed to review these theories/models and rethink their adequacy for Africa, Asia, Latin America, and the Caribbean. A new communications framework for HIV/AIDS was developed to move from a focus on the individual to a focus on five domains of "contexts" that influence behaviors: government policy, socioeconomic status (SES), culture, gender relations, and spirituality.


Government-owned inventions; availability for licensing. National Institutes of Health, Public Health Service, DHHS. Notice.
The invention listed below is owned by an agency of the U.S. Government and is available for licensing in the U.S. in accordance with 35 U.S.C. 207 to achieve expeditious commercialization of results of federally funded research and development.


A potent, nonpeptidyl 1H-quinolone antagonist for the gonadotropin-releasing hormone receptor.
Extensive development of the structure-activity relationships of a screening lead determined three important pharmacophores for gonadotropin-releasing hormone (GnRH) receptor antagonist activity. Incorporation of the 3,4,5-trimethylphenyl group at the 3-position, 2-(2(S)-azetidinyl)ethoxy group at the 4-position, and N-4-pyrimidinylcarboxamide at the 6-position of the quinolone core resulted in the identification of 4-(2-(azetidin-2(S)-yl)ethoxy)-7-chloro-2-oxo-3-(3,4,5-trimethylphenyl)-1,2-dihydroquinoline-6-carboxylic acid pyrimidin-4-ylamide (1) as a potent antagonist of the GnRH receptor. A 10(4)-fold increase in in vitro binding affinity is observed for the GnRH receptor as compared to the initial screening lead. Compound 1 exhibits nanomolar binding activity and functional antagonism at the human receptor and is 7-fold less active at the rhesus receptor. Intravenous administration of compound 1 to rhesus monkeys results in a significant decrease of the serum levels of downstream hormones, luteinizing hormone (79% decrease in area under the curve) and testosterone (92% decrease in area under the curve), at a dose of 3 mg/kg. Quinolone 1 is a potent nonpeptidyl antagonist for the human GnRH receptor that is efficacious for the suppression of luteinizing hormone and testosterone in primates.


Charcot-Marie-Tooth Disease and Other Genetic Polyneuropathies.
Genetic polyneuropathies are rare and clinically heterogeneous. This article provides an overview of the clinical features, neurologic and electrodiagnostic findings, and management strategies for Charcot-Marie-Tooth disease and other genetic polyneuropathies as well as an algorithm for genetic testing. In the past 10 years, many of the mutations causing genetic polyneuropathies have been identified. International collaborations have led to the development of consortiums that are undertaking careful genotype-phenotype correlations to facilitate the development of targeted therapies and validation of outcome measures for future clinical trials. Clinical trials are currently under way for some genetic polyneuropathies. Readers are provided a framework to recognize common presentations of various genetic polyneuropathies and a rationale for current diagnostic testing and management strategies in genetic polyneuropathies.


Purification and properties of pentachlorophenol hydroxylase, a flavoprotein from Flavobacterium sp. strain ATCC 39723.
A pentachlorophenol (PCP) hydroxylase which catalyzed the conversion of PCP to 2,3,5,6-tetrachlorohydroquinone and released iodide from triiodophenol in the presence of NADPH and oxygen was identified. The enzyme was purified by protamine sulfate precipitation, ammonium sulfate precipitation, hydrophobic chromatography, anion-exchange chromatography, gel filtration chromatography, and crystallization. The enzyme was a monomer with a molecular weight of 63,000. Under certain conditions, dimer and multimer conformations were also observed. The pI of the enzyme was pH 4.3. The optimal conditions for activity were a pH of 7.5 to 8.5 and a temperature of 40 degrees C. Each enzyme molecule contained one flavin adenine dinucleotide molecule. The Km for PCP was 30 microM and the Vmax was 16 mumol/min/mg of protein. The enzymatic reaction required 2 mol of NADPH per mol of halogenated substrate. On the basis of the data we present, it is likely that PCP hydroxylase is a flavoprotein monooxygenase. The addition of flavins to the reaction mixture did not stimulate the enzymatic reaction; however, we identified the photodegradation of triiodophenol and tribromophenol, but not PCP, by flavin mononucleotide or riboflavin and light.


Charge Transport in Sequence-Defined Conjugated Oligomers.
A major challenge in synthetic polymers lies in understanding how primary monomer sequence affects materials properties. In this work, we show that charge transport in single molecule junctions of conjugated oligomers critically depends on the primary sequence of monomers. A series of sequence-defined oligomers ranging from two to seven units was synthesized by an iterative approach based on the van Leusen reaction, providing conjugated oligomers with backbones consisting of para-linked phenylenes connected to oxazole, imidazole, or nitro-substituted pyrrole. The charge transport properties of these materials were characterized using a scanning tunneling microscope-break junction (STM-BJ) technique, thereby enabling direct measurement of molecular conductance for sequence-defined dimers, trimers, pentamers, and a heptamer. Our results show that oligomers with specific monomer sequences exhibit unexpected and distinct charge transport pathways that enhance molecular conductance more than 10-fold. A systematic analysis using monomer substitution patterns established that sequence-defined pentamers containing imidazole or pyrrole groups in specific locations provide molecular attachment points on the backbone to the gold electrodes, thereby giving rise to multiple conductance pathways. These findings reveal the subtle but important role of molecular structure including steric hindrance and directionality of heterocycles in determining charge transport in these molecular junctions. This work brings new understanding for designing molecular electronic components.


Low-risk lifestyle behaviors and all-cause mortality: findings from the National Health and Nutrition Examination Survey III Mortality Study.
We examined the relationship between 4 low-risk behaviors-never smoked, healthy diet, adequate physical activity, and moderate alcohol consumption-and mortality in a representative sample of people in the United States. We used data from 16958 participants aged 17 years and older in the National Health and Nutrition Examination Survey III Mortality Study from 1988 to 2006. The number of low-risk behaviors was inversely related to the risk for mortality. Compared with participants who had no low-risk behaviors, those who had all 4 experienced reduced all-cause mortality (adjusted hazard ratio [AHR]=0.37; 95% confidence interval [CI]=0.28, 0.49), mortality from malignant neoplasms (AHR=0.34; 95% CI=0.20, 0.56), major cardiovascular disease (AHR=0.35; 95% CI=0.24, 0.50), and other causes (AHR=0.43; 95% CI=0.25, 0.74). The rate advancement periods, representing the equivalent risk from a certain number of years of chronological age, for participants who had all 4 high-risk behaviors compared with those who had none were 11.1 years for all-cause mortality, 14.4 years for malignant neoplasms, 9.9 years for major cardiovascular disease, and 10.6 years for other causes. Low-risk lifestyle factors exert a powerful and beneficial effect on mortality.


[Management of diabetes in cardiac diseases].
Type 2 diabetes mellitus (DM) is a common disease affecting people in the world and its incidence is increasing rapidly. Cardiovascular disease (CVD) is the most important cause of mortality and morbidity among patients with type 2 DM. Patients with diabetes have a poorer prognosis than patients without diabetes. It is well known that other cardiovascular risk factors such as hyperlipidemia, hypertension and obesity that usually accompany to diabetes mellitus or impaired glucose tolerance as a metabolic syndrome component or not, can contribute to accelerated atherosclerosis and increased risk of cardiovascular event in diabetes mellitus. The management of patient with type 2 DM is not only glucocentric but focuses on multiple risk factor intervention.


Mineral status of COPD patients under long-term inhaled corticosteroid therapy.
The aim of this study was to determine the mineral status of mandibles, femurs, and spines in chronic obstructive pulmonary disease (COPD) patients under long-term inhaled corticosteroid therapy. Pulmonary function tests were conducted on patients (n = 30) with COPD under inhaled corticosteroid therapy for at least 1 year. The results were compared to sex- and age-matched controls (n = 30). Analyses of blood gases were also carried out relative to COPD, and bone mineral densities (BMD) of the mandible, lumbar spine, femoral neck, trochanter, and Ward's triangle were also measured by dual-energy X-ray absorptiometry (DEXA). Levels of serum osteocalcin, alkaline phosphatase, calcium, phosphorus, and cortisol were also assessed. In accordance with the Global Initiative for Chronic Obstructive Lung Disease criteria, 8 of the COPD patients had moderate, 11 patients had severe, and 11 patients had very severe forms of the disease. All BMD measurements were lower in the COPD patients than in the control group. The serum osteocalcin levels in COPD patients were significantly lower than those in the control group (p < 0.0001). Serum calcium (p < 0.004) and cortisol levels (p < 0.026) in the COPD patients were also significantly lower than those in the control subjects. Although serum alkaline phosphatase level was higher and the phosphorus level was lower in the treatment group than in the control group, the differences were not statistically significant. Regular evaluation of the biochemical markers of bone metabolism and BMD would be helpful for detecting any detrimental changes of bone in COPD patients under long-term inhaled corticosteroid therapy. In this study, mandibular BMD was observed to be lower in COPD patients under long-term inhaled corticosteroid therapy than in healthy subjects. Thus, dental implant treatment may require preventive measures in COPD patients under long-term inhaled corticosteroid therapy.


Click-Functionalization of a Poly(Tetrazine-co-Fluorene)-Conjugated Polymer with a Series of trans-Cyclooctene Derivatives.
A soluble poly(tetrazine) polymer was prepared via Suzuki polycondensation of 3,6-bis(5-bromofuran-2-yl)-1,2,4,5-tetrazine and a fluorene diboronate derivative. It can undergo efficient and quantitative post-polymerization inverse-electron-demand Diels-Alder click reactions with a variety of trans-cyclooctene (TCO) derivatives. The resulting polymers were oxidized to convert dihydropyridazine rings into pyridazines. The absorption spectra of the product polymers, both before and after oxidation, showed hypsochromic shifts that correlated with steric hindrance of the appended side chains. They also exhibited a significantly enhanced fluorescence intensity relative to the original poly(tetrazine). While gel-permeation chromatography indicated that the product polymers exhibited longer retention times, NMR end-group analysis showed that the polymers retained relatively constant degrees of polymerization. Graft copolymers were easily prepared via reaction with TCO-functionalized poly(ethylene glycol) chains and a cross-linked foam was produced by reacting the poly(tetrazine) with a bis-TCO crosslinker.


Amiodarone prophylaxis for atrial fibrillation after cardiac surgery: meta-analysis of dose response and timing of initiation.
To investigate a possible dose-response relationship between amiodarone and reduction in incidence of postoperative atrial fibrillation, and to determine whether pre- or postoperative initiation of amiodarone is superior. Meta-analysis of randomized controlled trials. MEDLINE and EMBASE databases and the Cochrane Central Register of Controlled Trials for English-language reports published between 1966 and December 2005. Of 23 identified randomized controlled trials of amiodarone prophylaxis of postoperative atrial fibrillation, 14 were included in the final analysis. These studies enrolled a total of 2864 patients. For each study, the total administered amiodarone dose--categorized as low (< 3000 mg), medium (3000-5000 mg), or high (> 5000 mg)--and preoperative versus postoperative initiation were aggregated by using meta-analytic techniques. The incidence of postoperative atrial fibrillation was significantly reduced by amiodarone compared with placebo (p<0.001). Although the odds of developing atrial fibrillation appeared to be somewhat higher in the low-dose group, no significant differences were noted in the odds ratios (ORs) of developing atrial fibrillation among the low-, medium-, and high-dose groups: OR 0.58, 95% confidence interval (CI) 0.44-0.77; OR 0.45, 95% CI 0.30-0.69; and OR 0.44, 95% CI 0.33-0.58; respectively (p=0.238). In addition, the ORs for atrial fibrillation development associated with preoperative and postoperative initiation of amiodarone were not significantly different (OR 0.50, 95% CI 0.39-0.63; and OR 0.48, 95% CI 0.37-0.63; respectively, p=0.862). Total amiodarone doses of 3000 mg or higher may be more effective than lower doses in reducing the rate of postoperative atrial fibrillation after cardiac surgery. Preoperative initiation of amiodarone appears to be unnecessary. These findings require confirmation in prospective, randomized trials.


miR-378 inhibits cell growth and enhances apoptosis in human myelodysplastic syndromes.
miR-378 has been proven to inhibit cell growth, migration and invasion in different types of cancers. In this study, we found that miR-378 was commonly downregulated in the bone marrow cells obtained from myelodysplastic syndrome (MDS) patients. We further investigated the role of miR-378 in the proliferation and apoptosis of SKM-1 cells, an acute myeloid leukemia cell line established in the leukemic phase during the progression of MDS to AML (MDS/AML). Results indicated that overexpression of miR-378 in SKM-1 cells interfered with proliferation via inducing apoptosis and G0/G1-phase cell cycle arrest, and suppressive effect of miR-378 on MDS/AML cells may be mediated partly through Bcl-w and CDC40. Moreover, apoptosis induced by miR-378 correlated with increased expression of Bax and activation of caspase-3, -8 and -9. Taken together, our data support a critical role for miR-378 in the pathogenesis of MDS and provide a novel therapeutic target in this complex disease.


Prognostic significance of haemodynamic parameters in patients with cardiogenic shock.
Invasive haemodynamic assessment with a pulmonary artery catheter is often used to guide the management of patients with cardiogenic shock (CS) and may provide important prognostic information. We aimed to assess prognostic associations and relationships to end-organ dysfunction of presenting haemodynamic parameters in CS. The Critical Care Cardiology Trials Network is an investigator-initiated multicenter registry of cardiac intensive care units (CICUs) in North America coordinated by the TIMI Study Group. Patients with CS (2018-2022) who underwent invasive haemodynamic assessment within 24 h of CICU admission were included. Associations of haemodynamic parameters with in-hospital mortality were assessed using logistic regression, and associations with presenting serum lactate were assessed using least squares means regression. Sensitivity analyses were performed excluding patients on temporary mechanical circulatory support and adjusted for vasoactive-inotropic score. Among the 3603 admissions with CS, 1473 had haemodynamic data collected within 24 h of CICU admission. The median cardiac index was 1.9 (25th-75th percentile, 1.6-2.4) L/min/m2 and mean arterial pressure (MAP) was 74 (66-86) mmHg. Parameters associated with mortality included low MAP, low systolic blood pressure, low systemic vascular resistance, elevated right atrial pressure (RAP), elevated RAP/pulmonary capillary wedge pressure ratio, and low pulmonary artery pulsatility index. These associations were generally consistent when controlling for the intensity of background pharmacologic and mechanical haemodynamic support. These parameters were also associated with higher presenting serum lactate. In a contemporary CS population, presenting haemodynamic parameters reflecting decreased systemic arterial tone and right ventricular dysfunction are associated with adverse outcomes and systemic hypoperfusion.


Electron density measurements in shock tube using microwave interferometry.
Microwave interferometry (MWI) is a nonintrusive diagnostic technique, capable of measuring small quantities of electrons present in a flame plasma. In this paper, a 94 GHz microwave interferometer is characterized and validated to perform robust and reliable measurements of electron concentrations in thermal and nonthermal plasmas in a shock tube. The MWI system is validated first by measuring the refractive index of a dielectric material. Subsequently, the system is used for measuring electron densities during the thermal ionization of argon and krypton in shock tube experiments. The measured activation energies are in good agreement with both the measured values from previous studies and theoretical values. The MWI system is finally used for measuring electron density time-histories in fuel oxidation experiments in the shock tube. The electron density profile of methane combustion shows a peak at the ignition time which agrees with pressure measurements. Experimental electron histories are also in overall agreement with predictions of the methane ion chemistry model.


Accuracy of actuarial procedures for assessment of sexual offender recidivism risk may vary across ethnicity.
Little is known about whether the accuracy of tools for assessment of sexual offender recidivism risk holds across ethnic minority offenders. I investigated the predictive validity across ethnicity for the RRASOR and the Static-99 actuarial risk assessment procedures in a national cohort of all adult male sex offenders released from prison in Sweden 1993-1997. Subjects ordered out of Sweden upon release from prison were excluded and remaining subjects (N = 1303) divided into three subgroups based on citizenship. Eighty-three percent of the subjects were of Nordic ethnicity, and non-Nordic citizens were either of non-Nordic European (n = 49, hereafter called European) or African Asian descent (n = 128). The two tools were equally accurate among Nordic and European sexual offenders for the prediction of any sexual and any violent nonsexual recidivism. In contrast, neither measure could differentiate African Asian sexual or violent recidivists from nonrecidivists. Compared to European offenders, AfricanAsian offenders had more often sexually victimized a nonrelative or stranger, had higher Static-99 scores, were younger, more often single, and more often homeless. The results require replication, but suggest that the promising predictive validity seen with some risk assessment tools may not generalize across offender ethnicity or migration status. More speculatively, different risk factors or causal chains might be involved in the development or persistence of offending among minority or immigrant sexual abusers.


Differential modulation of nitric oxide production by curcumin in host macrophages and NK cells.
Curcumin, the yellow pigment from Curcuma longa, has been shown to possess tumoricidal activity. We have earlier reported the induction of apoptosis in AK-5, rat histiocytic cells by curcumin leading to the inhibition of tumor growth in vivo. In this study we have observed differential activation status in host macrophages and NK cells induced by curcumin during the spontaneous regression of subcutaneously transplanted AK-5 tumors. Closer scrutiny of the cytokine profile and nitric oxide (NO) production by immune cells showed an initial downregulation of Th1 cytokine response and NO production by macrophages, and their upregulation in NK cells, which picked-up upon prolonged treatment with curcumin, culminating in a stronger tumoricidal effect. These studies suggest that the host macrophages and NK cells play an important modulatory role in the remission of AK-5 tumor.


Immunofluorescent and biochemical studies of terminal deoxynucleotidyl transferase in treated acute leukaemia.
Indirect Immunofluorescence (IF) for terminal deoxynucleotidyl transferase (TdT) was used in conjunction with the biochemical assay of TdT enzymatic activity to study human leukaemias before and during therapy. In addition, non-leukaemic marrows were analysed to compare the enzyme expression on normal cells. An excellent correlation was observed between the IF and biochemical methods when specimens contained greater than 5% TdT+ cells (by IF); below this level the biochemical assay was less reliable, while the sensitive IF test could detect isolated TdT+ cells among greater than 10 000 TdT negative cells. The IF method also had the advantage of allowing further immunological characterization of TdT+ cells, by simultaneous labelling of membrane antigens with appropriate antisera. TdT+ cells expressing Ia-like antigens (but lacking other antigens associated with B- and T- lymphoid differentiation) were frequently found in low numbers in remission marrows from acute lymphoblastic leukaemia (ALL) patients. However, similar cells were also observed in remission acute myeloid leukaemia, as well as in non-leukaemic regenerating marrows, and marrow from normal donors. The presence of these normal TdT+ precursor cells therefore precluded the use of either IF or biochemical TdT tests for estimating the degree of residual disease or predicting early relapse in patients with non-T, non-B ALL. In contrast, the detection of TdT+ cells with T lymphoid antigens (HuTLA+) but lacking Ia antigens, in thymic (T-cell)-ALL, but not in normal marrow, allowed the use of this combination of markers to detect minimal residual disease in T-ALL.


Butyric acid induces apoptosis by up-regulating Bax expression via stimulation of the c-Jun N-terminal kinase/activation protein-1 pathway in human colon cancer cells.
The colonic epithelial cells near the top of the crypt have been shown to undergo apoptosis. Because butyric acid (BA) is the major short-chain fatty acid produced by fermentation of dietary fiber in the large bowel, it may be an important regulator of apoptosis in colorectal cancer. We investigated which signaling pathway is triggered by BA to undergo apoptosis in human colorectal cancer cells. Human DiFi and FET colorectal cells were treated with BA to undergo apoptosis and were assayed for activation of c-Jun N-terminal kinase (JNK), transcription factor activation protein 1 (AP1) and NF-kappaB, and the proapoptotic molecule Bax. The contribution of specific pathways was assessed by examining the effects of dominant-negative mutants of JNK/AP1 or NF-kappaB on BA-induced Bax expression and apoptosis. BA-mediated DNA fragmentation and Bax induction were preceded by early stimulation of JNK, and the DNA-binding activities of AP1 and NF-kappaB. BA-induced enhancement of DNA fragmentation and stimulation of Bax promoter activity were blocked by the expression of dominant-negative mutants of JNK1 or AP1 but not NF-kappaB. These findings suggest that apoptosis triggered by BA involves transcriptional stimulation of the Bax gene via activation of the JNK/AP1 pathway in colonic epithelial cells.


The Subcostal Nerve During Lateral Approaches to the Lumbar Spine: An Anatomical Study with Relevance for Injury Avoidance and Postoperative Complications Such as Abdominal Wall Hernia.
Lateral approaches to the spine are increasing in popularity. However, details of the innervation pattern of the abdominal oblique muscles with the initial dissection have not been well studied. Ten adult fresh-frozen cadavers (20 sides) were placed in the lateral position. On each side, the region in which transpsoas approaches are performed, between the iliac crest and the 12th rib, was dissected. The nerves, their course, and their muscular supply were studied. The subcostal nerve is the predominant nerve supply for the anterolateral abdominal muscle innervation. It is larger and has a wider field of distribution and more branches (8 on average) compared with the L1 (4 on average) and 11th intercostal nerves (2 on average 2). The proximal 6-10 cm of each nerve has few if any branches. The subcostal nerve is often (75%) located up to 5 cm inferior to the 12th rib in its initial course. The area of least concentration ("safe zone") is located at an approximate midpoint between the lower edge of the 12th rib and the superior-most aspect of the iliac crest. A previously undescribed branch of the subcostal nerve was found traveling posterior to the quadratus lumborum and joining the remaining subcostal nerve in an anastomosis at or near the lateral position. Knowledge of the innervation and nerve dominance patterns might help decrease postoperative complications such as sensory deficits or abdominal wall hernias. The subcostal nerve is the dominant nerve in both size and innervation of the oblique muscles in the lateral position, transpsoas approach.


[The role of assessing UGT1A1 gene polymorphism in the prediction of irinotecan-induced toxicity in the course of chemotherapy for colorectal cancer].
There was performed a molecular genetic study of UGTlAl gene allelic variants polymorphism in patients with colorectal cancer who had had chemotherapy irinotecan-containing regimens FOLFIRI. Comparison of toxicity and the results of polymorphism of UGTlAl showed that dose-limiting hematologic and non-hematologic toxicities in patients with moderate and high risk of toxicity were higher (p = 0.050- 0.061) and the frequency of thrombocytopenia (p = 0.0257) and hyperbilirubinemia (p = 0.0439) were significantly higher compared to the low-risk group. Molecular genetic study of a complex examination of patients, which was planned to irinotecan should be performed to select the optimal dose and reduce the risk of toxicity of chemotherapy.


Extracorporeal photopheresis therapy in the management of steroid-refractory or steroid-dependent cutaneous chronic graft-versus-host disease after allogeneic stem cell transplantation: feasibility and results.
We conducted a retrospective analysis of all allogeneic stem cell transplantation (ASCT) patients started on extracorporeal photopheresis (ECP) for the management of steroid-dependent (SD) or steroid-refractory (SR) cutaneous chronic graft-versus-host disease (cGVHD) following ASCT during a 36-month period (9/98-8/01). Only SD or SR patients who were treated by ECP after day 100 and who received at least 4 weeks of ECP were considered evaluable for this analysis. Out of 64 ASCT patients reviewed, 32 patients met the inclusion criteria. All 32 patients had been previously treated with systemic corticosteroids with 11 (34%) being SR and 21 (66%) SD. Cutaneous cGVHD was extensive in 28 patients (88%) and was accompanied by visceral (hepatic, gastrointestinal) cGVHD in 23 patients (72%). The 32 evaluated patients had received a median of three prior therapies before ECP, most commonly systemic corticosteroids, tacrolimus, and mycophenolate mofetil. Patients received a median of 36 ECP sessions (range 12-98) over a median of 5.3 months (range 1-28), with a median of six sessions per month. The complete response (CR) rate was 22% (n=7) and the partial response rate was 34% (n=11). In all, 28 patients were on systemic corticosteroid therapy at ECP initiation and 18 patients achieved 50% dose reduction while on ECP, yielding a 64% steroid-sparing response rate. Of seven CRs, five are ongoing. A total of 11 (34%) patients have died after ECP, with all cases due to visceral cGVHD or cGVHD-related infectious complications. All 21 surviving patients remain on at least some immunosuppressive cGVHD therapy (including ECP in eight). Overall, ECP displays a substantial response rate and, in particular, steroid-sparing activity in SR/SD extensive cutaneous cGVHD. However, most patients continue to require at least some chronic therapy and cGVHD-related morbidity and mortality remain high.


Drug development and the importance of ethnicity: lessons from heart failure management and implications for hypertension.
Heart failure is a common condition, associated with both poor prognosis and poor quality of life. In contrast to all other cardiovascular diseases, the prevalence of heart failure is increasing in the western world, and is likely to continue to do so as the population ages. In the UK, a significant proportion of patients with heart failure come from South Asian and African Caribbean ethnic groups. A large body of evidence exists that there may be epidemiological and pathophysiological differences between patients with heart failure from different ethnic groups. Treatments such as ACE inhibitors, which are now part of standard heart failure therapy, have an evidence base consisting of trials in patients of almost exclusively white ethnicity. Such treatments may not be equally effective in patients from other ethnic groups. This review will discuss the current evidence for heart failure management with respect to ethnicity, and consider the implications for future drug development and implications for antihypertensive therapy.


Noncoding RNA as therapeutic targets for hepatocellular carcinoma.
Recent studies have suggested that noncoding RNAs (ncRNAs) contribute to the pathogenesis and progression of hepatocellular carcinoma (HCC). These RNA genes may be involved in various pathobiological processes such as cell proliferation, apoptosis, angiogenesis, invasion, and metastasis. Aberrant expression of ncRNA resulting from deregulated epigenetic, transcriptional, or posttranscriptional activity has been described in several studies. ncRNAs are comprised of a highly diverse group of transcripts that include microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) as well as several other types of RNA genes. Understanding the molecular mechanisms by which ncRNA contribute to hepatocarcinogenesis may enable the design of ncRNA-based therapeutics for HCC. In this review, the authors provide a perspective on therapeutic applications based on the emerging evidence of a contributory role of miRNAs and lncRNAs to the pathogenesis and progression of HCC. In addition, ncRNAs that are deregulated in expression in HCC may have utility as potential prognostic or diagnostic markers.


Ultrasound-guided pneumothorax induction prior to local anaesthetic thoracoscopy.
Local anaesthetic thoracoscopy (LAT) is performed by a growing number of respiratory physicians in the context of an expanding population with pleural disease. Most LATs occur in patients with moderate to large effusions where the presence of fluid allows safe access to the pleural space. Patients with little or no fluid, but other features concerning for pleural disease, are usually investigated by surgical means. Advanced LAT practitioners can also provide this service through pneumothorax induction, although there is little published data on the safety or efficacy of this technique. We present data from a series of 77 consecutive patients in whom ultrasound-guided pneumothorax induction and LAT were attempted. 67 procedures (87.0%) were successful, with the most common histopathological diagnoses being chronic pleuritis (58.2%) and mesothelioma (16.4%). No adverse events were reported secondary to the procedure. These findings demonstrate the utility of this approach and should inform future practice and guidelines.


Antibacterial and antioxidant properties of mixed linkage beta-oligosaccharides from extracted β-glucan hydrolysed by Penicillium occitanis EGL lichenase.
The aim of this study was first to ascertain the chemical composition and the physicochemical properties of cereal extracted β-glucan from barley flour. Secondly, to assess the antioxidant properties and the antibacterial properties of extracted β-glucan hydrolysates. The proximate composition, FT-IR and scanning electron microscopy of extracted β-Glucan were studied. Hydrolysates from extracted β-glucan, obtained by lichenase EGL from Penicillium occitanis, were a mixed linkage beta-oligosaccharides (MLBO) of trisaccharides and tetrasaccharides. MLBO showed a DPPH radical scavenger with IC50 about 1.8 ± 0.01 mg/mL whereas the IC50 of extracted β-glucan was about 5 ± 0.01 mg/mL. MLBO showed a high antioxidative capacity (175 μmol/mL α-tocopherol equivalents) at 5 mg/mL. The antimicrobial activity was confirmed against all tested bacteria especially at 20 mg/mL of MLBO while no inhibition was observed for all the strains used after the addition of either EGL or extracted β-glucan.


Mpv17l protects against mitochondrial oxidative stress and apoptosis by activation of Omi/HtrA2 protease.
Cellular localization determines whether the serine protease HtrA2 exerts pro- or antiapoptotic functions. In contrast to the well-characterized proapoptotic function of cytosolic HtrA2, mechanisms underlying the mitochondrial protective role are poorly understood. Mpv17l is a transmembrane protein previously implicated in peroxisomal reactive oxygen species metabolism and a close homolog of the inner mitochondrial membrane protein Mpv17. Here we demonstrate a previously undescribed direct interaction between Mpv17l and HtrA2 in mitochondria. The interaction is mediated by a PDZ domain and induces protease activation of HtrA2. HtrA2 inhibits mitochondrial superoxide generation, stabilizes mitochondrial membrane potential, and prevents apoptosis at baseline and in response to extracellular inducers of mitochondrial stress. The physiological role of Mpv17l is underscored by the finding that oxidative stress-induced downregulation of Mpv17l is a consistent feature in renal injury models. Our findings identify Mpv17l as a unique interacting protein and regulator of HtrA2 protease mediating antioxidant and antiapoptotic function in mitochondria.


Characterization of arylsulfatase C isozymes from human liver and placenta.
Arylsulfatase C and steroid sulfatase were thought to be identical enzymes. However, recent evidence showed that human arylsulfatase C consists of two isozymes, s and f. In this study, the biochemical properties of the s form partially purified from human placenta were compared with those of the f form from human liver. Only the placental s form has steroid sulfatase activity and hydrolyses estrone sulfate, dehydroepiandrosterone sulfate and cholesterol sulfate. The liver f form has barely detectable activity towards these sterol sulfates. With the artificial substrate, 4-methylumbelliferyl sulfate, both forms demonstrated a similar KM but the liver enzyme has a pH optimum of 6.9 while the placental form displayed two optima at 7.3 and 5.5. The molecular weight of the native enzyme determined with gel filtration was 183,000 for the s form and 200,000 for the f form and their pI's were also similar at 6.5. However, the T50, temperature at which half of the enzyme activity was lost, was 49.5 degrees C for the f form and 56.8 degrees C for the s form. Polyclonal antibodies raised against the placental form reacted specifically against the s and not the f form. They immuno-precipitated concomitantly greater than 80% of the total placental arylsulfatase C and steroid sulfatase activities while less than 20% of the liver enzyme was immuno-precipitable. In conclusion, the two isozymes s and f of arylsulfatase C in humans purified from placenta and liver, respectively, have similar KM, pI' and native molecular weight. However, they are distinct proteins with different substrate specificity, pH optima, heat-lability and antigenic properties. Only the s form is confirmed to be steroid sulfatase.


When the Hospital Is No Longer an Option: A Multiple Case Study of Defining Moments for Women Choosing Home Birth in High-Risk Pregnancies in The Netherlands.
Some women in a high-risk pregnancy go against medical advice and choose to birth at home with a "holistic" midwife. In this exploratory multiple case study, grounded theory and triangulation were employed to examine 10 cases. The women, their partners, and (regular and holistic) health care professionals were interviewed in an attempt to determine whether there was a pattern to their experiences. Two propositions emerged. The dominant one was a trajectory of trauma, self-education, concern about paternalism, and conflict leading to a negative choice for holistic care. The rival proposition was a path of trust and positive choice for holistic care without conflict. We discuss these two propositions and make suggestions for professionals for building a trusting relationship using continuity of care, true shared decision making, and an alternative risk discourse to achieve the goal of making women perceive the hospital as safe again.


Monochromatic phototherapy in elderly patients: a new way of treating chronic pressure ulcers?
Monochromatic pulsating light may have effects of importance for wound healing, and promising results were obtained in an open study on the healing of pressure ulcers, which motivated the present study. One hundred and sixty-four in- and outpatients with grade II and III ulcers were treated with monochromatic pulsating light or placebo over the ulcerated area, according to a specific therapeutic program. No significant effects were observed in the total material. In a subanalysis of grade II ulcers, there was a tendency toward better healing in the monochromatic light group (p = 0.06). A significantly larger reduction in pressure ulcer size was noted among patients in the treatment group with low body weight (BMI < 20). Monochromatic pulsating light may have effects on pressure ulcer healing, and a new study focusing only on grade II ulcers is under way.


Corpus callosum atrophy and neuropsychological outcome following carbon monoxide poisoning.
This study assessed the effects of carbon monoxide (CO) poisoning on the corpus callosum (CC). Sixty-two CO-poisoned patients had MRI scans and a battery of neuropsychological tests within 24 h (day of exposure) of CO poisoning and at 6 months post CO exposure. Serial quantitative magnetic resonance imaging (QMRI) analysis of the CC was carried out, with the day of exposure scans compared to the 6-month scans. There was no difference between normal subjects' CC and the baseline scans in the CO-poisoned patients. We detected a 15-mm2 loss in the cross-sectional surface area of the CC between baseline and the 6-month follow-up scans. The effect on the CC was generalized atrophy rather than CC subregion-specific changes. Neuropsychological test results performed at baseline and at 6 months did not correlate with the level of CC atrophy. Independent of any CC effects, the patients exhibited impaired memory, attention, and executive functioning on baseline testing, with variable improvement in cognitive function at 6 months. Quantitative MRI analysis allows for the detection of subtle CC changes that may not otherwise be observed following CO poisoning. The long-term effects of CO on the brain have been historically underestimated; however, we found subtle but significant CC atrophy and cognitive impairments following CO poisoning.


Mechanisms underlying achievement of final head position.
The studies reported here are directed toward understanding some of the mechanisms whereby the central nervous system terminates a given phase in a motor sequence and maintains a newly acquired position. In particular, we investigated the extent to which the termination of a centrally initiated head movement in monkeys and the subsequent maintenance of posture depend on a readout of proprioceptive afferent input generated during the movement itself or are instead centrally programmed. We approached this question in two ways: first, using vestibulectomized, but otherwise intact monkeys, we applied load disturbances unexpectedly at the beginning and throughout centrally initiated head movements with the aim of provoking a proprioceptive response in all types of neck receptors and to observe the outcome of this stimulation on the head final position. In a second set of experiments, we interrupted the flow of afferent input by cutting cervical and upper thoracic dorsal roots and observed how the absence of proprioceptive feedback affects the achievement of final head position. The results indicated that the central pattern of neural impulses establishing final head position is preprogrammed and it is not reset by the afferent proprioceptive impulses generated during the intended movement. In addition, our findings are consistent with the view that final head position is an equilibrium point dependent on a number of factors, such as the firing rate and the recruitment of the alpha motoneurons, the length-tension properties of the muscles involved in posture, and passive elastic properties of external loads.


Ribosome profiling the cell cycle: lessons and challenges.
Understanding the causes and consequences of dynamic changes in the abundance and activity of cellular components during cell division is what most cell cycle studies are about. Here we focus on control of gene expression in the cell cycle at the level of translation. The advent of deep sequencing methodologies led to technologies that quantify the levels of all mRNAs that are bound by ribosomes and engaged in translation in the cell (Ingolia et al. Science 324:218-223, 2009). This approach has been applied recently to synchronous cell populations to find transcripts under translational control at different cell cycle phases (Blank et al. EMBO J 36:487-502, 2017; Stumpf et al. Mol Cell 52:574-582, 2013; Tanenbaum et al. Elife 4:e07957, 2015). These studies revealed new biology, but they also have limitations, pointing to challenges that need to be addressed in the future.


A comparison of methods of adjusting stillbirth and neonatal mortality rates for birthweight in hospital and geographical populations.
Birthweight-specific rates can be useful for summarising stillbirth and neonatal mortality in populations but, sometimes, a single summary measure is required to compare several relatively small subpopulations. However, any particular summary has its shortcomings, and various methods have been proposed. We wished to compare mortality between local authorities and between hospitals in the Thames regions and thus required a single summary measure for each subpopulation. It was not obvious in advance which summary to use or whether a single method would work well for both local authorities (a geographical unit) and hospitals. This study compared six methods of calculating a single summary, three using indirect standardisation to adjust (500 g bands, 10-percentile bands, 10 z-score bands) and three using regression to adjust (mean birthweight, proportion < 2500 g, proportion < 1500 g). The data used were 570 016 births in the Thames Regions, broken down into its 96 local authorities and 65 hospitals. To investigate how well each adjustment had performed, we calculated the rank correlation between the crude and various adjusted mortality rates and mean birthweight, proportion < 2500 g and proportion < 1500 g. This was done separately in the local authorities and hospitals. If a method of adjustment had worked very well, these correlations should be negligible. For the local authorities, adjustment for proportion < 1500 g gave the lowest correlations. Adjustment for mean birthweight and 500 g-band standardisation did not appear to work so well but gave moderately low correlations. For hospitals, 500 g standardisation gave the lowest correlations. Adjustment for mean birthweight and proportion < 2500 g worked only moderately well. Percentile and z-score adjustment did not work well for local authorities or hospitals. We conclude that several methods appear to work reasonably well for local authorities, whereas for hospitals, 500 g indirect standardisation worked best. Percentile and z-score standardisation did not work well in these subpopulations.


Inter-observer agreement using the Canadian Emergency Department Triage and Acuity Scale.
To determine the inter-observer agreement on triage assignment by first-time users with diverse training and background using the Canadian Emergency Department Triage and Acuity Scale (CTAS). Twenty emergency care providers (5 physicians, 5 nurses, 5 Basic Life Support paramedics and 5 Advanced Life Support paramedics) at a large urban teaching hospital participated in the study. Observers used the 5-level CTAS to independently assign triage levels for 42 case scenarios abstracted from actual emergency department patient presentations. Case scenarios consisted of vital signs, mode of arrival, presenting complaint and verbatim triage nursing notes. Participants were not given any specific training on the scale, although a detailed one-page summary was included with each questionnaire. Kappa values with quadratic weights were used to measure agreement for the study group as a whole and for each profession. For the 41 case scenarios analyzed, the overall agreement was significant (quadratic-weighted K = 0.77, 95% confidence interval, 0.76-0.78). For all observers, modal agreement within one triage level was 94.9%. Exact modal agreement was 63.4%. Agreement varied by triage level and was highest for Level I (most urgent). A reasonably high level of intra- and inter-professional agreement was also seen. Despite minimal experience with the CTAS, inter-observer agreement among emergency care providers with different backgrounds was significant.


Cutting edge: cardiac myosin activates innate immune responses through TLRs.
Autoimmune attack on the heart is linked to host immune responses against cardiac myosin, the most abundant protein in the heart. Although adaptive immunity is required for disease, little is known about innate immune mechanisms. In this study we report that human cardiac myosin (HCM) acted as an endogenous ligand to directly stimulate human TLRs 2 and 8 and to activate human monocytes to release proinflammatory cytokines. In addition, pathogenic epitopes of human cardiac myosin, the S2 fragment peptides S2-16 and S2-28, stimulated TLRs directly and activated human monocytes. Our data suggest that cardiac myosin and its pathogenic T cell epitopes may link innate and adaptive immunity in a novel mechanism that could promote chronic inflammation in the myocardium.


JAK-STAT-mediated chronic inflammation impairs cytotoxic T lymphocyte activation to decrease anti-PD-1 immunotherapy efficacy in pancreatic cancer.
Human pancreatic cancer does not respond to immune check point blockade immunotherapy. One key feature of pancreatic cancer is the association between its progression and chronic inflammation. Emerging evidence supports a key role for the JAK-STAT pathway in pancreatic cancer inflammation. We aimed at testing the hypothesis that sustained JAK-STAT signaling suppresses cytotoxic T lymphocyte (CTL) activation to counteract anti-PD-1 immunotherapy-induced CTL activity in pancreatic cancer. We show that human pancreatic carcinomas express high level of PD-L1 and exhibit low level of CTL infiltration. JAK-STAT inhibitor Ruxolitinib selectively inhibits STAT1 and STAT3 activation and increases CTL infiltration to induce a Tc1/Th1 immune response in the tumor microenvironment in an orthotopic pancreatic cancer mouse model. Ruxilitinib-mediated tumor suppressive efficacy diminishes in T-cell-deficient mice. Pancreatic tumor grows significantly faster in IFNγ-deficient mice. However, neutralizing IFNγ does not alter tumor growth but diminishes Ruxolitinib-induced tumor suppression in vivo, indicating that lymphocytes and IFNγ are essential for Ruxolitinib-induced host antitumor immune response. Both type I and type II interferons upregulate PD-L1 expression through the JAK-STAT signaling pathway in mouse pancreatic tumor cells. Tumor cells respond to activated T cells by activating STAT3. The inhibition of STAT3 downregulates immune suppressive cytokines production by tumor cells, resulting in increased T cell activation and effector function. Consequently, Ruxolitinib significantly improves the efficacy of anti-PD-1 immunotherapy. Our data demonstrate that Ruxolitinib is effective in the inhibition of systemic inflammation in the tumor microenvironment and therefore upregulates CTL infiltration and activation to overcome pancreatic cancer resistance to anti-PD-1 immunotherapy.


Observations on the distribution of Salmonella in a pig abattoir.
A survey of 2211 pigs was carried out in a pig abattoir. Salmonella was isolated from 7-0 per cent of carcase swabs and from 11.6 per cent of 2205 samples of large intestinal contents. Salmonella typhimurium DT104 or DT104B was found in 3.2 per cent of the large intestinal samples (27.7 per cent of the salmonella isolates found) and in 2.7 per cent of the carcase swabs (38.7 per cent of the salmonella isolates from carcases). Within individual batches of pigs, the rate of isolation of salmonella ranged from 0 to 71.4 per cent from intestinal contents, and from 0 to 100 per cent on carcases. In a smaller subsurvey, salmonella was found in 9.1 per cent of intestinal contents, 5.8 per cent of mesenteric lymph nodes and 3.8 per cent of tonsils, but in no spleen samples. The rates of isolation of salmonella from pigs from different producer groups ranged from 0 to 21.9 per cent with 0 to 88.2 per cent of farm units within groups showing evidence of infection. S typhimurium DT104 or DT104B was isolated from large intestinal samples from 16.9 per cent of farm units. The number of salmonellae found in positive pigs ranged from 1.0 to 10(5) cfu/g intestinal contents and from 1.0 to 10(2) cfu per 0.1 m2 dressed carcase surface. Sequential sampling during the slaughter process showed that 82.9 per cent of the carcases were contaminated after bleeding, a proportion which was reduced to 5.7 per cent after scalding; after singeing none were contaminated. Estimations of surface enterobacteriaceae revealed a similar pattern of reductions and resurgences of contamination to that observed with salmonella. The rate of isolation of salmonella from pigs which had been held in lairage overnight was less than that from pigs slaughtered within two to three hours of arrival.


Oral lactobacilli isolated from teenage orthodontic patients.
Orthodontic bands form retention areas which create favourable conditions for growth of lactobacilli. This study deals with the numbers and specific species of lactobacilli in the oral cavity of teenage orthodontic patients. High numbers of lactobacilli were obtained (log 6.398 +/- 0.761 cfu/ml) and the buffer capacity of saliva indicated a high caries risk amongst these individuals but the actual DMFT values were relatively low. Homofermentative lactobacilli predominated in both plaque and saliva and particularly L. salivarius and L. casei. These two lactobacilli species indicated that these patients are at a high caries risk. Several other homofermentative and heterofermentative lactobacilli were isolated which indicated the complexity of the oral lactobacilli community in saliva and plaque.


Evolution of Callosal and Cortical Lesions on MRI in Marchiafava-Bignami Disease.
We report on serial MRI findings of cerebral lesions in a 55-year-old man with severe Marchiafava-Bignami disease (MBD). The first MRI change on fluid-attenuated inversion recovery images was hyperintensity in the genu of the corpus callosum and in the frontoparietal cortex. Following this change, a splenial lesion appeared. The first MRI change in the genu of the corpus callosum was not associated with a change in diffusion on diffusion-weighted MRI imaging, suggesting a pathological change involving vasogenic edema. Development of cortical lesions in the initial stage confirms that cortical lesions result from the primary pathogenetic process induced by alcoholic intoxication and malnutrition in MBD.


[Inhaled insulin, new perspective for insulin therapy].
Since the discovery of insulin and its use in diabetes care, patients, physicians and nurses dream of another way of insulin administration than the subcutaneous injections actually used. Different types of insulin administration have been evaluated and, particularly, that using the pulmonary route. The use of this alternative method to deliver insulin may result in improved patient compliance, facilitate intensified therapies and avoid the delay of initiating insulin administration because patient's reluctance. The different insulin pulmonary delivering devices actually studied will be presented. Preliminary data comparing this way of administration and the subcutaneous injection of human regular insulin are good, but sufficient data comparing inhaled insulin with the new short-acting insulin analogues are not yet available. Among various difficulties of the pulmonary insulin delivery, the finding of an effective promoter, capable of increasing the bioavailability of insulin, is a crucial issue. The cost of such insulin administration might also be a problem. Finally, careful studies concerning the safety of this kind of administration, particularly potential long-term pulmonary toxicity, are mandatory. Nevertheless, inhaled insulin is an attractive topic in which most important pharmaceutical companies are currently involved.


Studies on vacuolar membrane microdomains isolated from Arabidopsis suspension-cultured cells: local distribution of vacuolar membrane proteins.
The local distribution of both the vacuolar-type proton ATPase (V-ATPase) and the vacuolar-type proton pyrophosphatase (V-PPase), the main vacuolar proton pumps, was investigated in intact vacuoles isolated from Arabidopsis suspension-cultured cells. Fluorescent immunostaining showed that V-PPase was distributed evenly on the vacuolar membrane (VM), but V-ATPase localized to specific regions of the VM. We hypothesize that there may be membrane microdomains on the VM. To confirm this hypothesis, we prepared detergent-resistant membranes (DRMs) from the VM in accordance with well established conventional methods. Analyses of fatty acid composition suggested that DRMs had more saturated fatty acids compared with the whole VM in phosphatidylcholine and phosphatidylethanolamine. In the proteomic analyses of both DRMs and detergent-soluble mebranes (DSMs), we confirmed the different local distributions of V-ATPase and V-PPase. The observations of DRMs with an electron microscope supported the existence of different areas on the VM. Moreover, it was observed using total internal reflection fluorescent microscopy (TIRFM) that proton pumps were frequently immobilized at specific sites on the VM. In the proteomic analyses, we also found that many other vacuolar membrane proteins are distributed differently in DRMs and DSMs. Based on the results of this study, we discuss the possibility that VM microdomains might contribute to vacuolar dynamics.


Indication for 'Over the scope' (OTS)-clip vs. covered self-expanding metal stent (cSEMS) is unequal in upper gastrointestinal leakage: results from a retrospective head-to-head comparison.
Intestinal perforation or leakage increases morbidity and mortality of surgical and endoscopic interventions. We identified criteria for use of full-covered, extractable self-expanding metal stents (cSEMS) vs. 'Over the scope'-clips (OTSC) for leak closure. Patients who underwent endoscopic treatment for postoperative leakage, endoscopic perforation, or spontaneous rupture of the upper gastrointestinal tract between 2006 and 2013 were identified at four tertiary endoscopic centers. Technical success, outcome (e.g. duration of hospitalization, in-hospital mortality), and complications were assessed and analyzed with respect to etiology, size and location of leakage. Of 106 patients (male: 75 (71%), female: 31 (29%); age (mean ± SD): 62.5 ± 1.3 years, 72 (69%) were treated by cSEMS and 34 (31%) by OTSC. For cSEMS vs. OTSC, mean treatment duration was 41.1 vs. 25 days, p<0.001, leakage size 10 (1-50) vs. 5 (1-30) mm (median (range)), and complications were observed in 68% vs. 8.8%, p<0.001, respectively. Clinical success for primary interventional treatment was observed in 29/72 (40%) vs. 24/34 (70%, p = 0.006), and clinical success at the end of follow-up was 46/72 (64%) vs. 29/34 (85%) for patients treated by cSEMS vs. OTSC; p = 0.04. OTSC is preferred in small-sized lesions and in perforation caused by endoscopic interventions, cSEMS in patients with concomitant local infection or abscess. cSEMS is associated with a higher frequency of complications. Therefore, OTSC might be preferred if technically feasible. Indication criteria for cSEMS vs. OTSC vary and might impede design of randomized studies.


An eco-epidemiological modeling approach to investigate dilution effect in two different tick-borne pathosystems.
Disease (re)emergence appears to be driven by biodiversity decline and environmental change. As a result, it is increasingly important to study host-pathogen interactions within the context of their ecology and evolution. The dilution effect is the concept that higher biodiversity decreases pathogen transmission. It has been observed especially in zoonotic vector-borne pathosystems, yet evidence against it has been found. In particular, it is still debated how the community (dis)assembly assumptions and the degree of generalism of vectors and pathogens affect the direction of the biodiversity-pathogen transmission relationship. The aim of this study was to use empirical data and mechanistic models to investigate dilution mechanisms in two rodent-tick-pathogen systems differing in their vector degree of generalism. A community was assembled to include ecological interactions that expand from purely additive to purely substitutive. Such systems are excellent candidates to analyze the link between vector ecology, community (dis)assembly dynamics, and pathogen transmission. To base our mechanistic models on empirical data, rodent live-trapping, including tick sampling, was conducted in Wales across two seasons for three consecutive years. We have developed a deterministic single-vector, multi-host compartmental model that includes ecological relationships with non-host species, uniquely integrating theoretical and observational approaches. To describe pathogen transmission across a gradient of community diversity, the model was populated with parameters describing five different scenarios differing in ecological complexity; each based around one of the pathosystems: Ixodes ricinus (generalist tick)-Borrelia burgdorferi and I. trianguliceps (small mammals specialist tick)-Babesia microti. The results suggested that community composition and interspecific dynamics affected pathogen transmission with different dilution outcomes depending on the vector degree of generalism. The model provides evidence that dilution and amplification effects are not mutually exclusive in the same community but depend on vector ecology and the epidemiological output considered (i.e., the "risk" of interest). In our scenarios, more functionally diverse communities resulted in fewer infectious rodents, supporting the dilution effect. In the pathosystem with generalist vector we identified a hump shaped relationship between diversity and infections in hosts, while for that characterized by specialist tick, this relationship was more complex and more dependent upon specific parameter values.


Purification and cloning of a broad substrate specificity human liver carboxylesterase that catalyzes the hydrolysis of cocaine and heroin.
A human liver carboxylesterase (hCE-2) that catalyzes the hydrolysis of the benzoyl group of cocaine and the acetyl groups of 4-methylumbelliferyl acetate, heroin, and 6-monoacetylmorphine was purified from human liver. The purified enzyme exhibited a single band on SDS-polyacrylamide gel electrophoresis with a subunit mass of approximately 60 kDa. The native enzyme was monomeric. The isoelectric point of hCE-2 was approximately 4.9. Treatment with endoglycosidase H caused an increase in electrophoretic mobility indicating that the liver carboxylesterase was a glycoprotein of the high mannose type. The complete cDNA nucleotide sequence was determined. The authenticity of the cDNA was confirmed by a perfect sequence match of 78 amino acids derived from the hCE-2 purified from human liver. The mature 533-amino acid enzyme encoded by this cDNA shared highest sequence identity with the rabbit liver carboxylesterase form 2 (73%) and the hamster liver carboxylesterase AT51p (67%). Carboxylesterases with high sequence identity to hCE-2 have not been reported in mouse and rat liver. hCE-2 exhibited different drug ester substrate specificity from the human liver carboxylesterase called hCE-1, which hydrolyzes the methyl ester of cocaine. hCE-2 had higher catalytic efficiencies for hydrolysis of 4-methylumbelliferyl acetate, heroin, and 6-monoacetylmorphine and greater inhibition by eserine than hCE-1. hCE-2 may play an important role in the degradation of cocaine and heroin in human tissues.


Severe Maternal Morbidity Cases in Israel in a High-Volume High-Resource Referral Center: A Retrospective Cohort Study.
Admission to an intensive care unit (ICU) is an objective marker of severe maternal morbidity (SMM). To determine the prevalence of obstetric ICU admissions in one medical center in Israel and to characterize this population. In this retrospective study the files of women coded for pregnancy, birth, or the perinatal period and admission to the ICU were pulled for data extraction (2005-2013). During the study period, 111 women were admitted to the ICU among 120,279 women who delivered babies (0.09%). Their average age was 30 ± 6 years, most were multigravida, a few had undergone fertility treatments, and only 27% had complicated previous pregnancies. Most pregnancies (71.2%) were uneventful prior to admission. ICU admissions were divided equally between direct (usually hemorrhage) and indirect (usually cardiac disease) obstetric causes. The indications for obstetrics ICU admission correlated with the proximate causes of maternal arrest observed worldwide. While obstetric hemorrhage is often unpredictable, deterioration of heart disease is foreseeable. Attention should be directed specifically toward improving the diagnosis and treatment of maternal heart disease during pregnancy in Israel.


A probabilistic approach to learn chromatin architecture and accurate inference of the NF-κB/RelA regulatory network using ChIP-Seq.
Using nuclear factor-κB (NF-κB) ChIP-Seq data, we present a framework for iterative learning of regulatory networks. For every possible transcription factor-binding site (TFBS)-putatively regulated gene pair, the relative distance and orientation are calculated to learn which TFBSs are most likely to regulate a given gene. Weighted TFBS contributions to putative gene regulation are integrated to derive an NF-κB gene network. A de novo motif enrichment analysis uncovers secondary TFBSs (AP1, SP1) at characteristic distances from NF-κB/RelA TFBSs. Comparison with experimental ENCODE ChIP-Seq data indicates that experimental TFBSs highly correlate with predicted sites. We observe that RelA-SP1-enriched promoters have distinct expression profiles from that of RelA-AP1 and are enriched in introns, CpG islands and DNase accessible sites. Sixteen novel NF-κB/RelA-regulated genes and TFBSs were experimentally validated, including TANK, a negative feedback gene whose expression is NF-κB/RelA dependent and requires a functional interaction with the AP1 TFBSs. Our probabilistic method yields more accurate NF-κB/RelA-regulated networks than a traditional, distance-based approach, confirmed by both analysis of gene expression and increased informativity of Genome Ontology annotations. Our analysis provides new insights into how co-occurring TFBSs and local chromatin context orchestrate activation of NF-κB/RelA sub-pathways differing in biological function and temporal expression patterns.


Isotropic events observed with a borehole array in the Chelungpu fault zone, Taiwan.
Shear failure is the dominant mode of earthquake-causing rock failure along faults. High fluid pressure can also potentially induce rock failure by opening cavities and cracks, but an active example of this process has not been directly observed in a fault zone. Using borehole array data collected along the low-stress Chelungpu fault zone, Taiwan, we observed several small seismic events (I-type events) in a fluid-rich permeable zone directly below the impermeable slip zone of the 1999 moment magnitude 7.6 Chi-Chi earthquake. Modeling of the events suggests an isotropic, nonshear source mechanism likely associated with natural hydraulic fractures. These seismic events may be associated with the formation of veins and other fluid features often observed in rocks surrounding fault zones and may be similar to artificially induced hydraulic fracturing.


Hierarchical bayesian models for regularisation in sequential learning
We show that a hierarchical Bayesian modeling approach allows us to perform regularization in sequential learning. We identify three inference levels within this hierarchy: model selection, parameter estimation, and noise estimation. In environments where data arrive sequentially, techniques such as cross validation to achieve regularization or model selection are not possible. The Bayesian approach, with extended Kalman filtering at the parameter estimation level, allows for regularization within a minimum variance framework. A multilayer perceptron is used to generate the extended Kalman filter nonlinear measurements mapping. We describe several algorithms at the noise estimation level that allow us to implement on-line regularization. We also show the theoretical links between adaptive noise estimation in extended Kalman filtering, multiple adaptive learning rates, and multiple smoothing regularization coefficients.


Review: on TRAIL for malignant glioma therapy?
Glioblastoma (GBM) is a devastating cancer with a median survival of around 15 months. Significant advances in treatment have not been achieved yet, even with a host of new therapeutics under investigation. Therefore, the quest for a cure for GBM remains as intense as ever. Of particular interest for GBM therapy is the selective induction of apoptosis using the pro-apoptotic tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). TRAIL signals apoptosis via its two agonistic receptors TRAIL-R1 and TRAIL-R2. TRAIL is normally present as homotrimeric transmembrane protein, but can also be processed into a soluble trimeric form (sTRAIL). Recombinant sTRAIL has strong tumouricidal activity towards GBM cells, with no or minimal toxicity towards normal human cells. Unfortunately, GBM is a very heterogeneous tumour, with multiple genetically aberrant clones within one tumour. Consequently, any single agent therapy is likely to be not effective enough. However, the anti-GBM activity of TRAIL can be synergistically enhanced by a variety of conventional and novel targeted therapies, making TRAIL an ideal candidate for combinatorial strategies. Here we will, after briefly detailing the biology of TRAIL/TRAIL receptor signalling, focus on the promises and pitfalls of recombinant TRAIL as a therapeutic agent alone and in combinatorial therapeutic approaches for GBM.


Tumor suppressor RARRES1- A novel regulator of fatty acid metabolism in epithelial cells.
Retinoic acid receptor responder 1 (RARRES1) is silenced in many cancers and is differentially expressed in metabolism associated diseases, such as hepatic steatosis, hyperinsulinemia and obesity. Here we report a novel function of RARRES1 in metabolic reprogramming of epithelial cells. Using non-targeted LC-MS, we discovered that RARRES1 depletion in epithelial cells caused a global increase in lipid synthesis. RARRES1-depleted cells rewire glucose metabolism by switching from aerobic glycolysis to glucose-dependent de novo lipogenesis (DNL). Treatment with fatty acid synthase (FASN) inhibitor, C75, reversed the effects of RARRES1 depletion. The increased DNL in RARRES1-depleted normal breast and prostate epithelial cells proved advantageous to the cells during starvation, as the increase in fatty acid availability lead to more oxidized fatty acids (FAO), which were used for mitochondrial respiration. Expression of RARRES1 in several common solid tumors is also contextually correlated with expression of fatty acid metabolism genes and fatty acid-regulated transcription factors. Pathway enrichment analysis led us to determine that RARRES1 is regulated by peroxisome proliferating activated receptor (PPAR) signaling. These findings open up a new avenue for metabolic reprogramming and identify RARRES1 as a potential target for cancers and other diseases with impaired fatty acid metabolism.


First two cases of severe multifocal infections caused by Klebsiella pneumoniae in Switzerland: characterization of an atypical non-K1/K2-serotype strain causing liver abscess and endocarditis.
We describe the first two multifocal invasive infections due to Klebsiella pneumoniae recently observed in Switzerland. Phenotypic (MIC assays and string test) and molecular analyses (PCR/Sequencing for bla, virulence factor genes and whole genome sequencing for one strain) were performed to characterize the causative K. pneumoniae isolates. Both K. pneumoniae isolates (Kp1 and Kp2) were pan-susceptible to antibiotics and produced narrow-spectrum SHV β-lactamases. However, only Kp1 was string test positive. Kp1 was of ST380 and caused liver abscess as well as pneumonia and orbital phlegmon in an Eritrean patient. It belonged to the hypervirulent capsular serotype K2 and harboured the classic virulence-associated rmpA and aerobactin genes, fulfilling both the clinical and microbiological definitions for an invasive K. pneumoniae syndrome. Kp2 was of ST1043 and caused both liver abscess and endocarditis in a Swiss patient. Moreover, it did not possess the classic virulence-associated genes. Whole genome sequencing identified less well-known virulence factors in Kp2 that might have contributed to its virulence. Among these there were genes important for intestinal colonization and/or invasion, such as genes involved in adhesion (e.g., fimABCD and mrkABCD), regulation of capsule polysaccharide biosynthesis (e.g., evgS-evgA), as well as iron uptake (iroN), energy conversion, and metabolism. This report confirms the continuous dissemination of hypervirulent K. pneumoniae strains among patients of non-Asian descent in Europe. Moreover, it highlights the genetic background of an atypical hypervirulent K. pneumoniae causing a severe invasive infection despite not possessing the classical virulence characteristics of hypermucoviscous strains.


Hyperbaric oxygen treatment in delayed post-hypoxic encephalopathy following inhalation of liquefied petroleum gas: a case report.
Delayed post-hypoxic encephalopathy can occur after an episode of anoxia or hypoxia. Symptoms include apathy, confusion, and neurological deficits. We describe a 47-year-old male patient who inhaled gas from a kitchen stove liquid petroleum gas cylinder. He was diagnosed with hypoxic ischaemic encephalopathy 12 hours after his emergency department admission. He received six sessions of hyperbaric oxygen treatment (HBOT) and was discharged in a healthy state after six days. Fifteen days later, he experienced weakness, loss of appetite, forgetfulness, depression, balance problems, and inability to perform self-care. One week later, he developed urinary and fecal incontinence and was diagnosed with post-hypoxic encephalopathy. After 45 days from the onset of symptoms, he was referred to the Underwater and Hyperbaric Medicine Department for HBOT. The patient exhibited poor self-care and slow speech rate, as well as ataxic gait and dysdiadochokinesia. Hyperbaric oxygen was administered for twenty-four sessions, which significantly improved the patient's neurological status with only hypoesthesia in the left hand remaining at the end of treatment. Hyperbaric oxygen has been reported as successful in treating some cases of delayed neurological sequelae following CO intoxication. It is possible that HBO therapy may also be effective in delayed post-hypoxic encephalopathy from other causes. This may be achieved through mechanisms such as transfer of functional mitochondria to the injury site, remyelination of damaged neurons, angiogenesis and neurogenesis, production of anti-inflammatory cytokines, and balancing of inflammatory and anti-inflammatory cytokines.


[Polyurethane vessels for microvascular surgical training to reduce animal use].
Systematic training of the manual skills is inevitable in learning microsurgery. Generally, first exercises are done on two-dimensional models, then the training continues on animals. With the growing ethical awareness, the obligation to protect animals and the stricter animal protection laws, realistic three-dimensional models have become necessary for training of microsurgery. However, the available alternatives all have certain disadvantages. We tested vessels made of polyurethane for microvascular surgical training and compared them to the available three-dimensional synthetic alternatives. Rose-coloured (arteries) or blue (veins), opaque vessels with a minimal wall thickness of 0.12 mm and a minimal internal diameter of 1 mm are used. To mimic the surgical access and the depth of the operative field, the vessels can be embedded in a synthetic box with or without a cap. The completed anastomosis is checked by injection of a coloured fluid. The consistency and the variable relation of the thickness of the wall to the internal diameter very closely reflect the biological situation. Even training on very fragile venous walls is possible in all manners. After completion of anastomosis the vessels can be opened longitudinally to check the patency of the anastomotic site. The described polyurethane vessels are very suitable for microsurgical training as a useful step between the two-dimensional model and the animal. The number of animals required for microsurgical training can clearly be reduced by the use of such synthetic polyurethane vessels.


Generation of human-induced pluripotent stem cells to model spinocerebellar ataxia type 2 in vitro.
Spinocerebellar ataxia type 2 (SCA2) is caused by triple nucleotide repeat (CAG) expansion in the coding region of the ATAXN2 gene on chromosome 12, which produces an elongated, toxic polyglutamine tract, leading to Purkinje cell loss. There is currently no effective therapy. One of the main obstacles that hampers therapeutic development is lack of an ideal disease model. In this study, we have generated and characterized SCA2-induced pluripotent stem (iPS) cell lines as an in vitro cell model. Dermal fibroblasts (FBs) were harvested from primary cultures of skin explants obtained from a SCA2 subject and a healthy subject. For reprogramming, hOct4, hSox2, hKlf4, and hc-Myc were transduced to passage-3 FBs by retroviral infection. Both SCA2 iPS and control iPS cells were successfully generated and showed typical stem cell growth patterns with normal karyotype. All iPS cell lines expressed stem cell markers and differentiated in vitro into cells from three embryonic germ layers. Upon in vitro neural differentiation, SCA2 iPS cells showed abnormality in neural rosette formation but successfully differentiated into neural stem cells (NSCs) and subsequent neural cells. SCA2 and normal FBs showed a comparable level of ataxin-2 expression; whereas SCA2 NSCs showed less ataxin-2 expression than normal NSCs and SCA2 FBs. Within the neural lineage, neurons had the most abundant expression of ataxin-2. Time-lapsed neural growth assay indicated terminally differentiated SCA2 neural cells were short-lived compared with control neural cells. The expanded CAG repeats of SCA2 were stable throughout reprogramming and neural differentiation. In conclusion, we have established the first disease-specific human SCA2 iPS cell line. These mutant iPS cells have the potential for neural differentiation. These differentiated neural cells harboring mutations are invaluable for the study of SCA2 pathogenesis and therapeutic drug development.


Magnetic resonance imaging appearance of the shoulder after subacromial injection with corticosteroids can mimic a rotator cuff tear.
Subacromial injections have been used to treat rotator cuff problems. Previous studies have noted the difficulty in performing accurate injections into this area. In addition, one must also question the effects that misplaced corticosteroids could have on the surrounding tissues. In this case, a 51-year-old woman presented with several weeks of left shoulder pain and was diagnosed with rotator cuff tendonitis. After a subacromial injection with betamethasone and lidocaine, the patient noted 3 weeks of near complete pain relief, followed by a return of her symptoms. A magnetic resonance imaging scan obtained 7 weeks after the injection showed a full-thickness tear of the supraspinatus tendon. Five weeks later, the patient underwent arthroscopic evaluation of the shoulder and subacromial decompression. The rotator cuff tendons were noted to be intact and normal in appearance. The patient eventually had full resolution of her symptoms. Six months postoperatively, she underwent a new scan that showed a normal supraspinatus tendon. Apparently, the subacromial injection penetrated the anterior half of the supraspinatus tendon, causing a transient effect and signal change. One should use caution in the interpretation of magnetic resonance imaging scans of the shoulder soon after the injection of corticosteroids.


Genotypic Variation in Trichomonas vaginalis Detected in South African Pregnant Women.
Trichomonas vaginalis is the causative agent of trichomoniasis. The genetic characterisation of T. vaginalis isolates reveals significant genetic diversity in this organism. Data on the prevalence of different genotypes of T. vaginalis in South African populations is lacking. This study investigated the diversity of T. vaginalis in a pregnant population in South Africa. In this study, 362 pregnant women from the King Edward VIII Hospital in Durban, South Africa, provided vaginal swabs to be tested for the presence of T. vaginalis. T. vaginalis was detected using the TaqMan assay using commercially available primers and probes specific for this protozoan (Pr04646256_s1). The actin gene from T. vaginalis was amplified with gene-specific primers. The actin amplicons were digested with HindII, MseI, and RsaI, and the banding patterns were compared across the three digests for assignment of genotypes. Phylogenetic analysis was conducted using MEGA. The prevalence of T. vaginalis in the study population was 12.9% (47/362). Genotype G was the most frequent genotype in our study population. Genotypes H and I were detected in one sample each. According to the multiple sequence alignments and phylogenetic analysis, a level of diversity was observed across and within genotypes. Four different single-nucleotide changes in the actin gene were detected. Sample TV358 (H genotype) contained a single amino acid substitution from glutamine to lysine. Sample TV184 (G genotype) contained a single amino acid substitution from glutamic acid to arginine. Sample TV357 (G genotype) contained two amino acid substitutions, arginine to leucine and glycine to aspartic acid. Three different genotypes were observed in the pregnant population. Diversity was observed across and within genotypes. The observed diversity can be challenging for future vaccine design and development of antigen-based rapid diagnostic tests for trichomoniasis.


Acute subdural hematoma from bridging vein rupture: a potential mechanism for growth.
Most acute subdural hematomas (ASDHs) develop after rupture of a bridging vein or veins. The anatomy of the bridging vein predisposes to its tearing within the border cell layer of the dura mater. Thus, the subdural hematoma actually forms within the dura. The hematoma grows by continued bleeding into the border cell layer. However, the venous pressure would not be expected to cause a large hematoma. Therefore, some type of mechanism must account for the hematoma's expansion. Cerebral venous pressure (CVP) has been demonstrated in animal models to be slightly higher than intracranial pressure (ICP), and CVP tracks the ICP as pressure variations occur. The elevation of CVP as the ICP increases is thought to result from an increase in outflow resistance of the terminal portion of the bridging veins. This probably results from a Starling resistor model or, less likely, from a muscular sphincter. A hypothesis is derived to explain the mechanism of ASDH enlargement. Tearing of one or more bridging veins causes these vessels to bleed into the dural border cell layer. Subsequent ICP elevation from the ASDH, cerebral swelling, or other cause results in elevation of the CVP by increased outflow resistance in the intact bridging veins. The increased ICP causes further bleeding into the hematoma cavity via the torn bridging veins. Thus, the ASDH enlarges via a positive feedback mechanism. Enlargement of an ASDH would cease as blood within the hematoma cavity coagulates. This would stop the dissection of the dural border cell layer, and pressure within the hematoma cavity would equalize with that in the torn bridging vein or veins.


Treponema pallidum does not synthesise in vitro a capsule containing glycosaminoglycans or proteoglycans.
Treponema pallidum was investigated for its ability to synthesise glycosaminoglycans or proteoglycans in vitro. Isolated viable T pallidum organisms were incubated with radiolabelled precursors of glycosaminoglycans, sodium 35S-sulphate and 3H-glucosamine (tritiated glucosamine). T pallidum failed to incorporate sodium 35S-sulphate but did incorporate 3H-glucosamine into a macromolecule which may be associated with the surface of the treponeme. This macromolecule was resistant to degradation by specific glycosaminoglycanases. We conclude that T pallidum does not synthesise a capsule containing glycosaminoglycans in vitro.


Regulation of blood L-glutamate levels by stress as a possible brain defense mechanism.
Isoflurane-anesthetized rats submitted to a closed head injury (CHI) display a significant decrease of their blood glutamate levels. Having demonstrated that a decrease of blood L-glutamate (glutamate) causes an increase of the driving force for a spontaneous brain-to-blood glutamate efflux, and consequently affords brain neuroprotection, we investigated here the possible mechanisms which can affect blood glutamate levels. Reasoning that the spontaneous decrease of blood glutamate levels post CHI could be part of a stress response, we observed that the stress involved in tail artery catheterization under isoflurane anesthesia does not affect blood glutamate levels. Investigating in naïve rats the stress effectors, we found that corticotropin-releasing factor (CRF) significantly decreased blood glutamate levels. Pretreatment with antalarmine (a selective type-1 CRF receptor antagonist) occludes the CRF-mediated decrease in blood glutamate levels. In contrast, the adrenocorticotrophic hormone (ACTH) did not affect blood glutamate levels. Investigating the effectors of the sympathetic/adrenomedullary system, we observed that in naïve rats, adrenaline but not noradrenaline decreased blood glutamate levels. Confirming the role of adrenaline, propranolol pretreatment (a non-selective beta-antagonist) prevented the spontaneous decrease of blood glutamate observed post CHI. On the strength of these results, we further observed that isoproterenol (a beta(1/2)-selective adrenoreceptor agonist) produced a marked sustained decrease in blood glutamate levels. These results suggest that stress induces a decrease of blood glutamate levels partly via the activation of peripheral CRF receptors and the activation of the beta-adrenoreceptors. We propose that this newly identified component of the stress response could be a peripherally mediated defense mechanism of the injured brain against the deleterious effects of excess glutamate.


Epidemiology of Sepsis-like Illness in Young Infants: Major Role of Enterovirus and Human Parechovirus.
Sepsis-like illness is a main cause for hospital admission in young infants. Our aim was to investigate incidence, epidemiology and clinical characteristics of enterovirus (EV) and human parechovirus (HPeV) infections in young infants with sepsis-like illness. This is a prospective observational cohort study in which infants younger than 90 days of age, presenting with sepsis-like symptoms in a secondary care children's hospital, underwent a full sepsis work-up. Clinical signs and infectious indices were recorded. EV or HPeV RNA was detected by polymerase chain reaction in plasma and/or cerebrospinal fluid (CSF). Infants were diagnosed with EV, HPeV, fever of unknown origin or severe infection. EV and HPeV were detected in 132 of 353 (37%) and 52 of 353 (15%) of cases, respectively. EV and HPeV have distinct seasonability. Some differences in clinical signs and symptoms occurred between children with EV and HPeV infection but were of limited clinical value. CSF pleocytosis occurred in 44% of EV positive infants, and only in 13% of those with HPeV infection. EV and HPeV infections are major causes of sepsis-like illness in infants < 90 days of age. Neither clinical characteristics nor laboratory indices were predictive for EV/HPeV infection. CSF pleocytosis occurs, but not in all patients. Testing for EV and HPeV in all young infants with sepsis-like illness is strongly advised.


The European Court legitimates access of Italian couples to assisted reproductive techniques and to pre-implantation genetic diagnosis.
On 28 August 2012, the European Court of Human Rights (ECHR) issued a judgment regarding the requirements for the legitimate access of couples to assisted reproductive techniques (ART) and to pre-implantation genetic diagnosis (PGD). This judgment concerns the case of an Italian couple who found out after their first child was born with cystic fibrosis that they were healthy carriers of the disease. When the woman became pregnant again in 2010 and underwent fetal screening, it was found that the unborn child also had cystic fibrosis, whereupon she had the pregnancy terminated on medical grounds. In order to have the embryo genetically screened prior to implantation under the procedure of PGD, the couple sought to use in vitro fertilisation to have another child. Since article 1 of the Italian law strictly limits access to ART to sterile/infertile couples or those in which the man has a sexually transmissible disease, the couple appealed to the European court, raising the question of the violation of articles 8 and 14 of the European Convention on Human Rights. The applicants lodged a complaint that they were not allowed legitimate access to ART and to PGD to select an embryo not affected by the disease. The European Court affirmed that the prohibition imposed by Italian law violated article 8 of the European Convention on Human Rights. Focusing on important regulatory and legal differences among EU Nations in providing ART treatments and PGD, we derived some important similarities and differences.


Chronic tolerance and sensitization to alcohol in sons of alcoholics: II. Replication and reanalysis.
Data from D. B. Newlin and J. B. Thomson (1991) were reanalyzed, and data from an independent replication study were analyzed, relative to tonic (baseline) and phasic (response to alcohol challenge) aspects of drinking alcohol administered at the same dose on several occasions. Among the high-risk men (sons of alcoholic fathers), linear trends across days for resting (predrinking) baselines were opposite to alcohol-evoked changes for finger pulse amplitude, finger temperature, and skin conductance in Study 1 and for pulse transit time and body sway (static ataxia) in Study 2. In contrast, the structure of the low-risk men's (sons of nonalcoholic parents) data was precisely the opposite. Results are discussed in terms of sensitization as a potential mechanism that relates vulnerability to final manifestation of addictive behavior.


Latex allergy: everyone is at risk.
Latex allergy often begins innocently with no symptoms or a slight case of contact dermatitis. As the allergy progresses, it becomes life-threatening. While this process puts our patients at great risk, we as health care workers fail to realize the risk we place our health and careers in, as we allow ourselves to become further exposed to the latex allergen at work and at home.


Cerebral injury after cardiac surgery: identification of a group at extraordinary risk. Multicenter Study of Perioperative Ischemia Research Group (McSPI) and the Ischemia Research Education Foundation (IREF) Investigators.
Cerebral injury after cardiac surgery is now recognized as a serious and costly healthcare problem mandating immediate attention. To effect solution, those subgroups of patients at greatest risk must be identified, thereby allowing efficient implementation of new clinical strategies. No such subgroup has been identified; however, patients undergoing intracardiac surgery are thought to be at high risk, but comprehensive data regarding specific risk, impact on cost, and discharge disposition are not available. We prospectively studied 273 patients enrolled from 24 diverse US medical centers, who were undergoing intracardiac and coronary artery surgery. Patient data were collected using standardized methods and included clinical, historical, specialized testing, neurological outcome and autopsy data, and measures of resource utilization. Adverse outcomes were defined a priori and determined after database closure by a blinded independent panel. Stepwise logistic regression models were developed to estimate the relative risks associated with clinical history and intraoperative and postoperative events. Adverse cerebral outcomes occurred in 16% of patients (43/273), being nearly equally divided between type I outcomes (8.4%; 5 cerebral deaths, 16 nonfatal strokes, and 2 new TIAs) and type II outcomes (7.3%; 17 new intellectual deterioration persisting at hospital discharge and 3 newly diagnosed seizures). Associated resource utilization was significantly increased--prolonging median intensive care unit stay from 3 days (no adverse cerebral outcome) to 8 days (type I; P<0.001) and from 3 to 6 days (type II; P<0.001), and increasing hospitalization by 50% (type II, P=0.04) to 100% (type I, P<0.001). Furthermore, specialized care after hospital discharge was frequently necessary in those with type I outcomes, in that only 31% returned home compared with 85% of patients without cerebral complications (P<0.001). Significant risk factors for type I outcomes related primarily to embolic phenomena, including proximal aortic atherosclerosis, intracardiac thrombus, and intermittent clamping of the aorta during surgery. For type II outcomes, risk factors again included proximal aortic atherosclerosis, as well as a preoperative history of endocarditis, alcohol abuse, perioperative dysrhythmia or poorly controlled hypertension, and the development of a low-output state after cardiopulmonary bypass. These prospective multicenter findings demonstrate that patients undergoing intracardiac surgery combined with coronary revascularization are at formidable risk, in that 1 in 6 will develop cerebral complications that are frequently costly and devastating. Thus, new strategies for perioperative management--including technical and pharmacological interventions--are now mandated for this subgroup of cardiac surgery patients.